Protective effect of glutamine pretreatment on HK-2 cells injury induced by anoxiareoxygenation

Objective To determine whether glutamine(GLN) pretreatment may protect human proximal renal tubular epithelial(HK-2) cells against anoxia-reoxygenation injury and the role of Heme oxygenase1(HO-1). Methods Cells were randomly assigned to four groups: control group(Con) : in which cells were untreated; CoCl2 group(CoCl2) : in which cells were treated with 300 μM CoCl2(chemical anoxia analogue) for 1 hour, cultured with normal medium for 24 hours, and then stimulated with no serum media; GLN group: in which cells were pretreated with 10 mmol /L GLN for 24 hours, then treated by the same method as group CoCl2; ZnPP group(inhibitor of HO-1) : cells were treated with 3 μM ZnPP and 10 mmol /L GLN for 24 hours, then treated by the same method as group CoCl2. Using MTT method and flow cytometry to detect the proliferation and apoptosis of HK-2 cells,RT-PCR method was used to investigate the regulation of HO-1,NF-κB,Bcl-2 and caspase-3 mRNA. Results After pretreated with 10 mmol /L GLN, the HK-2 cells activity was increased and the apoptosis was decreased(P 0. 01), the expression of HO-1 and Bcl-2 gene were up-regulated,NF-κB and caspase-3 gene were down-regulated(P 0. 05 or 0. 01). Conclusion Pretreatment with 10 mmol /L GLN effectively prevents the HK-2 cells against anoxia-reoxygenation injury by regulating the expression of apoptosis related genes,HO-1 induced by GLN plays a key role in this cytoprotection.